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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 10, Iss. 25 — Dec. 16, 2002
  • pp: 1458–1464

Fluorescence spectrum estimation using multiple color images and minimum negativity constraint

Maricor Soriano, Wilma Oblefias, and Caesar Saloma  »View Author Affiliations

Optics Express, Vol. 10, Issue 25, pp. 1458-1464 (2002)

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An inexpensive method to convert a microscope into an imaging spectrometer is presented. Unlike current microscope-based spectrometers which use specialized optics or scanning mechanisms, our system only requires at most two image captures with a 3-chip CCD camera and a lightly-tinted color filter to output the color signal of a sample at each pixel. Basis spectra are obtained by principal components analysis applied to an ensemble of color signals of commercially-available dyes observed with different dichroic mirrors. A transformation matrix from channel values to spectral coefficients is derived. Minimum negativity constraint is applied to eliminate negative parts of the reconstructed fluorescence spectrum. The technique is demonstrated on fluorescence microspheres (fluorospheres) and chlorophyll from plant leaf.

© 2002 Optical Society of America

OCIS Codes
(180.2520) Microscopy : Fluorescence microscopy
(300.0300) Spectroscopy : Spectroscopy
(330.1690) Vision, color, and visual optics : Color

ToC Category:
Research Papers

Original Manuscript: October 2, 2002
Revised Manuscript: November 28, 2002
Published: December 16, 2002

Maricor Soriano, Wilma Oblefias, and Caesar Saloma, "Fluorescence spectrum estimation using multiple color images and minimum negativity constraint," Opt. Express 10, 1458-1464 (2002)

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